EFFICACY OF TEST FORMULATIONS IN A RAT MODEL OF BENIGN PROSTATIC HYPERPLASIA

EFFICACY OF TEST FORMULATIONS IN A RAT MODEL OF BENIGN PROSTATIC HYPERPLASIA

 

1.0  TEST SYSTEM DETAILS:

Species                   : Rattus norvegicus (Rat)

Age                        : 10-12 weeks

Sex                        : Male

No. of animals        : 8 /Group

Total animals         : 48

 

2.0   ALLOCATION OF GROUPS:

 

Group No.	Group Description	Disease Induction agent administered	Treatment administered	Dose Volume and Route
G1	Normal Control	Treated with 0.1 ml of sesame oil × 28-conscutive days	0.5% MC, p.o., q.d.	5 ml/kg, p.o.
G2	Disease Control	Subcutaneous Injection with 2 mg/day of testosterone propionate mixed with estradiol benzoate at the ratio of 100:1 for 28-consecutive days.	0.5% MC, p.o., q.d..
G3	Reference Control		Finasteride 1 mg/kg, p.o., q.d. in 0.5% MC
G4	Treated orally with low dose of TF		TF -X1 mg/kg, in 0.5% MC., q.d.
G5	Treated orally with intermediate dose 1 of TF		TF -X2 mg/kg, in 0.5% MC., q.d.
G6	Treated orally with intermediate dose 2 of TF		TF -X3 mg/kg, in 0.5% MC., q.d.

 

Abbreviations: MC-Methyl Cellulose, p.o.-per os. q.d.: quaque die; X1, X2, and X3, are defined as the incremental doses of the Test formulations. The dose range will be from 10 mg/kg to 1000 mg/kg, q.d.

 

3.0  METHOD:

·         After completion of quarantine, healthy animals will be selected for the study. Subsequently, they will be randomized based on body weight and allocated into 6, different groups consisting of 8 animals each.

·         Post-randomization, animals will be acclimatized for 5 days in an experimental room earmarked for the experiment.

·         For the induction of benign prostate hyperplasia, animals allocated to groups G2-G6 will be subcutaneously injected with 2 mg/day of testosterone propionate mixed with estradiol benzoate at the ratio of 100:1 in sesame oil for 28-consecutive days. The daily dose of testosterone propionate and estradiol benzoate will be 2 mg and 0.02 mg, respectively. The animals assigned to the normal control group (G1) will be treated with 0.1 ml of sesame oil.

Ø Animals of Group G1 and G2, designated as normal-control and disease-control respectively, will be administered 0.5% MC, p.o., q.d.

Ø Animals of group G3 will be treated with the reference drug, Finasteride (1mg/kg), which will be suspended in 0.5% MC and will be given orally at the dose of 1 mg/kg, p.o., q.d.

Ø Animals of group G4-G6 will be treated with TF orally at different dose levels ranging from 10-1000 mg/kg, q.d.

·         On day 29, i.e. twenty-eight days of treatment of testosterone propionate and estradiol benzoate, animals will be sacrificed under an overdose of thiopentone anaesthesia. After suitable anaesthesia but before the animal dies, blood will be collected and serum will be separated for estimation of IL-8, TNF-a, DHT. Immediately after the animal dies, ventral prostatic lobes, seminal vesicles, and bladder will be harvested and weighed. One portion will be fixed in 10% neutral buffered formalin for histopathological analysis. Further, the remaining tissue will snap-frozen in liquid nitrogen and immediately stored at -80°C for the subsequent biochemical and molecular evaluations.

 

4.0  PARAMETERS TO BE EVALUATED:

·         Body weight: Once a week

·         Weight of the ventral prostatic lobes, seminal vesicles, and bladder.

·         Assessment of Prostate Weight (PW) and Prostatic Index (PI).

·         ELISA Assay of IL-8, Tumor Necrosis Factor (TNF)-a, and DHT in serum and prostate tissue.

·         Immunohistochemical Staining of Proliferating Cell Nuclear Antigen (PCNA).

·         Measurement of Superoxide Dismutase (SOD) Activity and Malondialdehyde (MDA) Concentration.

·         Immunofluorescent Assay of Caspase-3.

·         Histological analysis of all the ventral prostatic lobes, seminal vesicles, and bladder (Hematoxylin & Eosin-stained).

 

5.0  REFERENCES:

 

1.        Li, Z. et al. Upregulation of oxytocin receptor in the hyperplastic prostate. Front. Endocrinol. (Lausanne). 9, 1–13 (2018).

2.        Zhang, J. et al. Animal models of benign prostatic hyperplasia. Prostate Cancer Prostatic Dis. 24, 49–57 (2021).

3.        Cai, H., Zhang, G., Yan, Z. & Shang, X. The Effect of Xialiqi Capsule on Testosterone-Induced Benign Prostatic Hyperplasia in Rats. Evidence-based Complement. Altern. Med. 2018, (2018).

END OF DOCUMENT

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